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This volume explores a collection of methods that studies genome editing across a variety of bacteria, phages, and plants. Chapters in this book cover topics such as scarless DNA recombineering of phage in the lysogenic state; HEMSE; Dup-In and DIRex; recombineering in Staphylococcus aureus; and genome editing with Cas9 in lactobacilli. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and thorough, Recombineering: Methods and Protocols is a valuable resource for any researchers interested in learning more about this developing field.

This volume discusses the latest online plant genomics and cytogenetic resources used by plant evolutionary biologists and plant breeders. The chapters in this book are organized into two parts. Part One looks at plant genomic databases, and covers topics such as plant phenomics and genomics research data repositories, InpactorDB, PlanTEenrichment, and PEATmoss, among others. Part Two looks at cytogenetics and chromosome-related databases, and covers resources such as the Plant DNA C-values database, the Delphineae Chromosome Database (DCDB), B-chrom, a Database on B-chromosomes, and the Plant Ribosomal DNA Database. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective databases and offers explicit directions on how to access and get the most of these resources. Cutting-edge and comprehensive, Plant Genomic and Cytogenetic Databases is a valuable instrument for any plant science researcher who is interested in learning more about the wealth of information that is available through the use of these databases.

This detailed volume explores experimental laboratory procedures for a wide range of steroid bioconversions. After an overview on the current trends and perspectives, the book continues with sections covering microbial screening and synthetic biology applied to microorganisms able to catabolize sterols, methods on strain characterization, including omics and biochemical analyses, methods of fermentation and biocatalysis for steroids production, as well as a chapter on the medical use of glucocorticoids in cancer patients. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Microbial Steroids: Methods and Protocols, Second Edition serves as an ideal reference source for laboratory and industrial professionals, as well as for students in a wide array of biological disciplines.

This detailed volume provides the increasing number of SARS-CoV-2 researchers with a useful handbook covering multidisciplinary approaches on various aspects of SARS-CoV-2 research, brought together by leading laboratories across the globe. Topics covered include techniques in clinical and diagnostic virology, basic protocols in cell and virus culture, as well as bioinformatics and proteomics approaches in cellular response studies. This comprehensive collection also covers methods in immunology, animal models, antivirals and vaccine development strategies, as well as biorisk and mitigation measurements for SARS-CoV-2 research. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detailed implementation advice that is vital for success in the lab. Practical and timely, SARS-CoV-2: Methods and Protocols serves as an ideal guide for scientists investigating this prevalent and perilous RNA virus and the novel coronavirus disease that results from it.

This second edition details new and updated methods on different antibody libraries, along with novel approaches for antibody discovery. Chapters focuses on the construction of antibody libraries, antibody expression, complementary approaches for antibody selection, and other phage display related applications. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Phage Display: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.

This third edition volume expands on the previous editions with new discussions on the latest techniques and developments in the field. The chapters in this book are organized into four parts, and cover topics such as optical tweezers; single-molecule fluorescence tools; atomic force microscopy; magnetic tweezers; applications to virus protein shells, unfolding of proteins, nucleic acids, motor proteins, in vivo and in vitro; and protocols to establish specific surface interactions and perform force calibration. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and thorough, Single Molecule Analysis: Methods and Protocols, Third Edition is a valuable resource for all researchers who wantto learn more about this exciting and still expanding field.Chapters 2, 7, 8, 9, 12, 18, and 19 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This second edition details protocols that analyze and explore gene regulatory networks (GRNs). Chapters guide readers through experimental techniques used to study genes and their regulatory interactions in plants, and computational approaches used for the integration of experimental data and bioinformatics-based predictions of regulatory interactions. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Plant Gene Regulatory Networks: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.

This second volume focuses on PCR methods and PCR application specificities to the biotechnology and bioengineering field. New and updated chapters detail real-time PCR protocols, synthetic biology applications, pathogen detection, microfluidics, digital, multiplex detection recent advances. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, PCR: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.

This detailed book explores new molecular biological techniques for the detection of both antigens and antibodies of porcine diseases. Assays examined within this collection include improved methods of nucleic acid extraction, polymerase chain reaction (PCR), droplet digital PCR (ddPCR), polymerase spiral reaction (PSR), cross-priming amplification (CPA), enzyme-linked immunosorbent assay (ELISA), as well as peptide nucleic acid (PNA) based tools, aptamer-based tools, and lateral flow assays and different immune assay-based diagnostics for porcine diseases. As part of the Springer Protocols Handbooks series, the present book contains the type of in-depth knowledge on important diagnostics tools that provide a vital source of information for scientists, researchers, and students from different fields. Authoritative and practical, Protocols for the Diagnosis of Pig Viral Diseases aims to guide researchers toward developing new technologies for the diagnosis of pig pathogens.

This volume highlights natural products, molecular methods for identifying, and current trends in designing non-natural natural products. Chapters guide readers through protocols on heterologous expression techniques, gene disruption, modified pathway regulators, and in-vitro studies. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.Authoritative and cutting-edge, Engineering Natural Product Biosynthesis: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge. Chapter 13 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This second edition volume discusses the latest techniques and protocols used in the field that were not covered in the previous edition. The chapters in this book are organized into five parts. Part One looks at transcriptomic analyses and Part Two covers DNA replication and protein/DNA interactions. Part Three discusses translation dynamics, protein complexes, and proteomics. Part Four looks at genotypic screens and phenotypic profiling, and Part Five explores in silico integration of functional genomics data. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary material and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting edge and practical, Yeast Functional Genomics: Methods and Protocols, Second Edition is a valuable resource for all researchers interested in learning more about the evolving field of yeast.Chapters 1, 9, 16, 20, 22, 24, and 25 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This second edition provides detailed practical information on important methods employed in the engineering of plant secondary metabolism pathways. New and updated chapters guide readers through extraction, quantification, purification, localization, characterization, data mining and processing, biosynthesis modulation, and pathway engineering of representative classes of plant specialized metabolites. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, list of necessary materials and reagents, tips on troubleshooting and known pitfalls, and step-by-step descriptions of readily reproducible protocols. Authoritative and cutting-edge, Plant Secondary Metabolism Engineering: Methods and Protocols, Second Edition aims to be a useful practical guide to help researchers working in this exciting field.

This third edition details new and updated methods and protocols on important databases and data mining tools. Chapters guides readers through archives of macromolecular sequences and three-dimensional structures, databases of protein-protein interactions, methods for prediction conformational disorder, mutant thermodynamic stability, aggregation, and drug response. Quality of structural data and their release, soft mechanics applications in biology, and protein flexibility are considered, too, together with pan-genome analyses, rational drug combination screening and Omics Deep Mining. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials, includes step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Data Mining Techniques for the Life Sciences, Third Edition aims to be a practical guide to researches to help further their study in this field.

This volume explores the fields of stem cell biology, regenerative medicine, and cancer biology. The chapters in this book cover topics such as embryonic stem cells (ES) and induced pluripotent cells (iPS); ES/iPSC derived tissue stem cells; tissue resident stem cells; assays for primitive malignant cells; and CRISPR/Cas9 engineering and transgenic model development. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Stem Cell Assays: Methods and Protocols is a valuable laboratory manual that allows researchers and scientists to learn about new techniques and approaches in this developing field.

This collection explores state-of-the-art methods and protocols for research on photodynamic therapy (PDT) and its use in a wide range of medical applications, from antiviral to anticancer. Beginning with an extensive section on in vitro and in vivo models, the volume continues with chapters on oxygen-independent photosensitizers, next-generation photosensitization strategies, contemporary insights into the immunomodulatory effects of PDT, antimicrobial effects of PDT, as well as a variety of general biochemical and molecular biological techniques. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detailed implementation advice that ensures successful results in the lab. Thorough and authoritative, Photodynamic Therapy: Methods and Protocols serves as an ideal source of inspiration for both new and established PDT scientists and a guide for designing innovative research programs in this continuously advancing and multidisciplinary field.

This volume details features of DNA methylation data, data processing pipelines, quality control measures, data normalization, and to discussions of statistical methods for data analysis, control of confounding and batch effects, and identification of differentially methylated regions. Chapters focus on microarray-based methylation measures and sequence-based measures. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary methodologies and software packages, step-by-step, readily reproducible analysis pipelines, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Epigenome- Wide Association Studies: Methods and Protocols: aims to be a useful practical guide to researches to help further their study in this field.

This volume provides methods on the role of TLRs in immune responses and on the various approaches targeting different TLRs in cancer, autoimmunity and in vaccine development. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Toll-Like Receptors: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.

This book analyzes liquid biopsy applications in cancer and other diseases. Chapters guide readers through the latest technologies and analysis methods for liquid biopsy,liquid biopsy in cancer, role of liquid biopsies in rheumatoid arthriti, cell-free circulating DNA profiling in patients with skin diseases, circulating non-coding RNAs, and exomes. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Liquid Biopsies: Methods and Protocols aims to attract more researchers and clinicians to study the diagnosis, immunotherapy, and prognosis of cancer and other diseases with liquid biopsy analysis.

This volume explores updated and entirely new experimental approaches used to investigate phagocytosis and phagosome maturation. In order to aid in the study of engulfment, maturation, resolution, and pathogen manipulation of phagocytes, the book features methodology to quantify uptake and maturation specific to certain phagocytes, particles, or pathogens, while other chapters offer methods that can be applied generically across the field. Methods are presented to study phagocytosis and phagosome maturation in vivo, in cellulo, and through in vitro analyses. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Phagocytosis and Phagosomes: Methods and Protocols, Second Edition serves as a vital resource for both experts in the field as well as for investigators delving into phagocytosis and phagosome maturation for the first time.

This volume explores the conceptual framework and the practical issues related to genomic prediction of complex traits in human medicine and in animal and plant breeding. The book is organized into five parts. Part One reminds molecular genetics approaches intending to predict phenotypic variations. Part Two presents the principles of genomic prediction of complex traits, and reviews factors that affect its reliability. Part Three describes genomic prediction methods, including machine-learning approaches, accounting for different degree of biological complexity, and reviews the associated computer-packages. Part Four reports on emerging trends such as phenomic prediction and incorporation into genomic prediction models of "omics" data and crop growth models. Part Five is dedicated to lessons learned from cases studies in the fields of human health and animal and plant breeding, and to methods for analysis of the economic effectiveness of genomic prediction. Written in the highly successful Methods in Molecular Biology series format, the book provides theoretical bases and practical guidelines for an informed decision making of practitioners and identifies pertinent routes for further methodological researches. Cutting-edge and thorough, Complex Trait Predictions: Methods and Protocols is a valuable resource for scientists and researchers who are interested in learning more about this important and developing field.Chapters 3, 9, 13, 14, and 21 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This second edition details new and updated chapters on key methodologies and breakthroughs in the mass spectrometry imaging (MSI) field. Chapters guide readers through nano-Desorption Electrospray Ionisation (nDESI), Matrix Assisted Laser Desorption Ionisation-2 (MALDI-2), Laser Ablation - Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS) ,Imaging Mass Cytometry (IMC) with a variety of diverse samples including eye tissue, crop analysis, 3D cell culture models, and counterfeit goods analysis. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Imaging Mass Spectrometry: Methods and Protocols, Second Edition aims to be a useful and practical guide to new researchersand experts looking to expand their knowledge.

This volume looks at the latest methods used to study imaging techniques, metabolic tracing, and deep muscle phenotyping. The chapters in this book cover topics such as imaging skeletal muscle mass using MRI, dual-energy x-ray absorptiometry, CT, and ultrasound; measures of neuromuscular function such as power/strength/force; microscopy and immunohistochemistry; and ex vivo fibre function and isolation. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Comprehensive and thorough, Neuromuscular Assessments of Form and Function is a valuable resource for researchers interested in multiple methods used to study skeletal muscle neurophysiology.

This book aims to deliver the most up-to-date protocols in using epiblast stem cells (EpiSC) to answer critical questions on mammalian development. Beginning with a section on the spectrum of mouse primed pluripotent stem cells, the volume continues with chapters on toolkits to characterize and study primed stem cells, as well as primed stem cells to engineer gastrulation models. Written for the highly successful Method in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Epiblast Stem Cells: Methods and Protocols serves as an ideal guide to overcome the reproducibility problem by gathering detailed state-of-the-art protocols to perform comprehensive analyses, enabling the reader to opt for the appropriate cell type to model the epiblast.

This book discusses principles, methodology, and applications of microbiological laboratory techniques . It lays special emphasis on the use of various automated machines that are essential for medical microbiology and diagnostic labs. The book contains eleven major chapters. The first chapter describes the good lab practices which should be followed by the students in all biological, chemistry or microbiology laboratories. The next chapter describes manual and automated characterization of antibiotic resistant microbes, followed by a chapter on genomics based tools and techniques that are integral to research. Further chapters deal with other important techniques like immunology based techniques, spectrophotometry and its various types, MALDI-TOFF and microarrays, each with illustrations and detailed description of the protocols and applications. The book also gives certain important guidelines to the students about the planning the experiment and interpreting results.The book is highly informative and provides latest techniques. It is a handy compendium for graduate and post graduate students, as well as more advanced researchers.

This second edition describes up-to-date methodological approaches, ranging from physiological assays to imaging and molecular techniques, to study a wide variety of plant responses to environmental cues. Chapters are divided into four sections detailing protocols to investigate key biological processes underlying plant environmental responses in the model organisms Arabidopsis thaliana and Physcomitrella patens as well as in different crop species. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Environmental Responses in Plants, Second Edition aims to be a foundation for future studies and to be a source of inspirationfor new investigations in the field.

This detailed book provides an exhaustive picture of current methods to detect, isolate, and analyze extracellular vesicles (EVs) from diverse sources, now seen as potential disease biomarkers as well as tools for the development of new therapies. Beginning with a section on detection and isolation of EVs, the volume continues with chapters covering different methods to isolate and quantify EVs from specialized tissues/organs and body fluids, methods devoted to analyzing EV components, as well as cutting-edge methods to engineer EVs. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Extracellular Vesicles in Diagnosis and Therapy serves as an ideal guide for researchers seeking to learn more about the complexities of cell-to-cell communication.

This detailed volume explores a wide variety of applications of yeast surface display, an extensively used protein engineering technology. Beginning with detailed protocols for the construction and efficient selection/screening of yeast surface display libraries, as well as for the analysis of individual yeast-displayed protein variants, the book continues with protocols describing the selection of yeast surface display libraries for binding to mammalian cells or to extracellular matrix as well as protocols for a broad spectrum of specialized yeast surface display applications, demonstrating the versatility of this display platform. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible methodologies, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Yeast Surface Display serves as a comprehensive resource that enables the implementation of this powerful and versatile technique in virtually any molecular biology laboratory, even in the absence of any prior yeast surface display experience.

This volume provides state-of-the-art methodology to functionalize microtubules and construct microtubule-based materials. Chapters consists of six sections detailing modification of microtubule structures, observation and control of microtubule movement, material applications of microtubules in vitro, development of microtubule-binding molecules, comprehensive approaches to analyze properties of microtubules, and functionalization of microtubules in living cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Microtubules: Methods and Protocols aims to provide useful information for construction of microtubule-based materials and help to create new ideas for the next-generation of materials and applications through detailed protocols.

This volume provides protocols and methods on techniques to study plant gametogenesis. Chapters are divided into four sections covering omics, cytological, molecular approaches, plant transformation, genome editing, bioinformatics, and data analysis. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Plant Gametogenesis: Methods and Protocols aims to be a foundation for future studies and to be a source of inspiration for new investigations in the field.

This volume details the inventory and characterizes traditional meat products within each geographic region. Consisting of 21 chapters, chapters guide readers through comprehensive descriptions of the materials and processing conditions used to make each meat product so that it can be manufactured by other researchers or industries. In addition, each book chapter includes explanatory notes and elucidate the possible specific points to take into account for the correct manufacture. Authoritative and cutting-edge, Production of Traditional Mediterranean Meat Products aims to standardize the manufacturing process of the main Mediterranean traditional meat products, since there are several variations between manufacturers or regions.